International Journal of Chemical Studies 2020; 8(5): 2393-2397
P-ISSN: 2349–8528
E-ISSN: 2321–4902
www.chemijournal.com
IJCS 2020; 8(5): 2393-2397
© 2020 IJCS
Received: 08-07-2020
Accepted: 12-08-2020
Jules Yoda
1. Department of Traditional Medicine
and Pharmacopeia-Pharmacy
(MEPHATRA/Ph)/ Research
Institute for Health Sciences (IRSS),
Ouagadougou, Burkina Faso
2. Laboratory of Molecular Chemistry
and Materials (LC2M) /Team of
Organic Chemistry and
Phytochemistry (ECOP) / University
Joseph Ki-Zerbo, Ouagadougou,
Burkina Faso
Benjamin Ouédraogo
Laboratory of Environmental Analytical
Chemistry and Bio Organics
(LCAEBiO), University Joseph KiZerbo, Ouagadougou, Burkina Faso
Sosso Siaka
Laboratory of Molecular Chemistry and
Materials (LC2M) /Team of Organic
Chemistry and Phytochemistry (ECOP) /
University Joseph Ki-Zerbo,
Ouagadougou, Burkina Faso
Félix B Kini
1. Department of Traditional Medicine
and Pharmacopeia-Pharmacy
(MEPHATRA/Ph)/ Research
Institute for Health Sciences (IRSS),
Ouagadougou, Burkina Faso
2. Laboratory of Environmental
Analytical Chemistry and Bio
Organics (LCAEBiO), University
Joseph Ki-Zerbo, Ouagadougou,
Burkina Faso
Abdoulaye Djandé
Laboratory of Molecular Chemistry and
Materials (LC2M) /Team of Organic
Chemistry and Phytochemistry (ECOP) /
University Joseph Ki-Zerbo,
Ouagadougou, Burkina Faso
Adama Saba
Laboratory of Molecular Chemistry and
Materials (LC2M) /Team of Organic
Chemistry and Phytochemistry (ECOP) /
University Joseph Ki-Zerbo,
Ouagadougou, Burkina Faso
Corresponding Author:
Jules Yoda
1. Department of Traditional Medicine
and Pharmacopeia-Pharmacy
(MEPHATRA/Ph)/ Research
Institute for Health Sciences (IRSS),
Ouagadougou, Burkina Faso
2. Laboratory of Molecular Chemistry
and Materials (LC2M) /Team of
Organic Chemistry and
Phytochemistry (ECOP) / University
Joseph Ki-Zerbo, Ouagadougou,
Burkina Faso
Phytochemical investigation of five herbal teas
useful in mental disorders treatment
Jules Yoda, Benjamin Ouédraogo, Sosso Siaka, Félix B Kini, Abdoulaye
Djandé and Adama Saba
DOI: https://doi.org/10.22271/chemi.2020.v8.i5ag.10676
Abstract
Mental disorders or illnesses are common and affect all countries. According to official estimates, in
most countries at least one person in three is affected. Several researches have shown the benefits of
plants in the management of these pathologies. This study focuses on five medicinal plants useful in the
management of mental disorders, including Cymbopogon citratus, Lippia chevalieri, Combretum
micratum, Chrysantellum americanum and Tetrapleura tetraptera. These plants are used in Burkina Faso
in the form of herbal teas. The objective of this study is to enhance the value of these plants and improve
their use by some physico-chemical analyses, a phytochemical study and the determination of antioxidant
activity. Among the parameters that will be precisely investigated there are organoleptic analysis,
moisture content, extraction yields, thin layer chromatographic fingerprinting, phytochemical screening
and the determination of total phenol, total flavonoids. The DPPH method was used to evaluate the
antioxidant power of the various aqueous extracts. The physico-chemical analysis gave satisfactory
results on the main indices. The phytochemical study allowed to establish the chromatographic
fingerprints of the aqueous extracts, to identify the main bioactive secondary metabolites and to
determine the content of total phenols and total flavonoids in the extracts of the five plants. Interesting
results are obtained, which confirms the use of these plants. Among the samples submitted to the
different analyses, the extract of Combretum micratum presented high levels of total phenolics (395,38 ±
7,05 mg GAE/g extract) and total flavonoids (105,87 ± 0,05 mg Ru E/g extract). This extract also
exhibited the highest antiradical activity.
Keywords: Mental disorders, herbal teas, phytochemical screening, phenolic compounds, antiradical
properties
1. Introduction
Mental disorder or mental illness is a behavioural pattern that results in impaired personal
functioning [1]. Common mental disorders generally refer to two broad categories of diagnoses
including depressive disorders and anxiety disorders. These conditions are among the most
common causes of morbidity and disability worldwide [2, 3]. According to estimates, the most
common mental disorders are depression, which affects about 264 million people, bipolar
disorder, which affects about 45 million people, dementia, which affects about 50 million
people, and schizophrenia and other psychoses, which affect about 20 million people
worldwide [4]. The causes of these diseases are often multiple and confusing, which makes the
management by modern medicine difficult. In some countries, phytotherapy is found as a safe
alternative in the treatment of several mental illnesses [5]. Despite the great advances in modern
medicine in recent years, especially in the field of organic synthesis of bioactive molecules,
plants still contribute significantly to the improvement of the health of the world population,
especially in underdeveloped countries [6, 8]. Plants, with their wide variety of chemical
constituents, offer a promising source of medicines for many diseases, including mental
disorders. This present study concerns Cymbopogon citratus, Lippia chevalieri, Combretum
micratum, Chrysantellum americanum and Tetrapleura tetraptera involved in the treatment of
mental illness [9, 13]. These five plants have been used as herbal teas for years in some West
African countries, i.e. Burkina Faso. The objective of this study is to contribute to the
valorization of these medicinal plants through chemical investigations such as
chromatographic fingerprinting, phytochemical screening and phenolic compound content.
Antioxidant activity was also evaluated using antiradical test with DPPH reagent.
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2. Materials and Methods
2.1. Sample Collection and Preparation
The plant material consisted of organs of the five species
including the leaves of Cymbopogon citratus, Combretum
micranthum, Lippia chevalieri, the whole plant of
Chrysantellum americanum and the the fruits of Tetrapleura
tetraptera. These plant parts were collected in october 2019 in
two locations. They were dried away from light, moisture and
dust. They are then pulverized by means of a blade mill and
the powders thus obtained will be used to carry out the
various extraction operations.
2.2. Organoleptic Tests
The examination of organoleptic characteristics includes
analyses of the general shape, colour, smell of the drug [14].
2.3. Moisture Content
A test sample is weighed and the resulting mass is recorded.
The drying process is repeated until the moisture mass of 1g
of powder sample was placed a in the crucible and dried in an
oven at 105 °C. After cooling, the steamed content attained a
constant value [15].
2.4. Extraction Yield
The extracts used for phytochemical screening are obtained
by depletion of the different crude aqueous extracts with
solvents in order of increasing polarity (n-hexane,
dichloromethane, ethylacetate, butanol) [14]. For the other
analyses, the extracts were obtained by aqueous decoction.
2.4. Fingerprinting by Thin Layer Chromatography
Each sample was solubilized (10 mg/mL) and 5 μL was
deposited on a 60 F254 silica gel plate (glass support). The
chromatographic plate is then immersed in the tank containing
the desired mobile phase for the development of the
chromatogram over a distance of 8 cm. After the
chromatogram has been developed, the chromatographic plate
is visualized using a UV lamp. The TLC was conducted
according to the general methods of the Pharmacopoeia.
2.5. Determination of the Phytochemical Constituent
The phytochemical screening of the plants extracts was
carried out by simple qualitative methods [16].
2.6. Total Phenolics Content
Total phenolic content were determined by Singleton method
[17]
. These compounds react with the Folin Ciocalteu reagent
(FCR) in an alkaline medium. The loss of a phenolic proton in
an alkaline medium leads to a phenolate anion that is capable
to reducing. the FCR in molybdate forming a blue colored
molybdenum oxide complex with a maximum absorption at
760 nm. The intensity of the blue coloration is proportional to
the amount of total phenolics present in the test sample. The
reaction mixture consisted of 1 ml extract, 1 ml 2N FCR and
3 ml 20% sodium carbonate solution. It was left at rest at
room temperature for 40 min and then the absorbance is
measured at 760 nm on the spectrophotometer (Agilent 8453).
In the white control tube, the extract was replaced by distilled
water. A standard curve will be drawn with tannic acid (1-5
µg/ml).
2.7. Total Flavonoids Content
The flavonoid assay was performed using the Kumaran
method [18] adapted by Abdel-Hameed [19]. 2 ml of extract of
concentration 1 mg/ml in methanol were mixed with 2 ml of
2% aluminium trichloride in methanol. After 40 minutes, the
absorbance was measured at 415 nm using the
spectrophotometer (Agilent 8453). The white control tube
consisted of 2 ml of methanol and a standard curve was
plotted with quercetin. The tests were carried out in triplicate.
2.8. Determination of Antiradical Properties
The technique used to determine the antiradical activity of
extracts is based on the methodology developed by Takao
with slight modifications [20]. 1,1-diphenyl-2-picrylhydrazyl
(DPPH), the reference oxidant, is dissolved in methanol at a
concentration of 2 mg /mL and used to spray TLC plates after
migration.
3. Resultats and Discussion
3.1. Organoleptic test and moisture content
To obtain the organoleptic characteristics, we determined the
main indices such as color, odor and flavor of the raw powder
of each of the five plants. The results are recorded in Table 1.
Table 1: Organoleptic characteristics and moisture content
Crude powder
colour
odour
Cymbopogon citratus
Lippia chevalieri
Combretum micratum
Chrysantellum americanum
Tetrapleura tetraptera
greenish
greenish
greenish
greenish
brown
citratus scent
odourless
citratus scent
pleasant scent
honey scent
Moisture
Content (%)
7,39
6,99
3,94
3,01
3,84
The powders each have an odor but without a characteristic
taste. These tests, although preliminary, sometimes make it
possible to immediately recognize the plant drug, to check its
degree of purity according to the presence or absence of
foreign elements, moulds, etc. And possibly to detect
adulteration or falsification [8, 14]. It is also possible to propose
polyherbal teas by an association of 2 or 3 of these powders
by exploiting its different indices.
The water loss of the samples ranged from 3.84% ± 0.11 to
7.39% ± 0.08 (Table1). An excessively high percentage of
water in a drug promotes a number of enzymatic reactions
that can negatively affect the appearance of the drug, its
organoleptic characteristics, and its therapeutic properties by
degrading the active ingredients over time [14]. Thus, the
content allowed in a medicinal product for a good
conservation should not exceed 10% [21]. Thus, the moisture
content of all our powders being less than 10%, this suggests
that our herbal medicines could be stored for a long time with
less risk of contamination and/or alteration of chemical
principles.
3.2. Extraction and Chromatographic Fingerprint
The extracts a-d are obtained by depleting the different
powders with solvents of increasing polarity. The extract e by
aqueous decoction. The extraction yield obtained is given in
the table below (Table 2). Extracts a (Hexanic extracts);
Extracts b (dichloromethane extracts); Extracts c (Ethyl
acetate extracts); Extracts d (butanolic extracts); Extracts e
(aqueous decoction.)
Table 2: The extraction yield
Extraxts
(Organic and Aqueous)
Cymbopogon citratus
Lippia chevalieri
Combretum micratum
Chrysantellum americanum
Tetrapleura tetraptera
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Extrac Extracts Extracts Extracts Extracts
ts a
b
c
d
e
1,51
2,27
3,43
4,64 23,375
2,57
2,50
3,90
4,88
16,96
1,67
2,05
4,25
5,86
16,34
1,05
3,40
21,20
8,82
22,53
10,00 2,93
2,61
5,86
39,46
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The lowest yields are obtained with hexane and the highest
with butanol and aqueous decoction. The yield of organic
butanol and aqueous decoated extracts is related to the
presence of polar compounds such as polyphenols.
Regarding the developed chromatograms of the aqueous
extract observed under the UV lamp at 366 nm, the plate was
sprayed with a specific flavonoid reagent several distinct
spots can be observed. The 5 chromatograms contain blue,
yellow, brown, orange, green spots attesting the presence of
flavonoids. The Tetrapleura tetraptera contains fewer spots
and those found are blue in color. In all the samples we note
the presence of phenolic compounds. A phytochemical
screening would allow us to detect other phenolic compounds
involved in the management of mental illness.
(1): Cymbopogon citratus; (2): Chrysantellum americanum; (3): Combretum
micratum (4): Lippia chevalieri; (5): Tetrapleura tetraptera
Fig 1: Fingerprint of aqueous extracts
3.3 Phytochemical Screening
The various extracts were analysed by TLC in order to
highlight the chemical interest groups.
The results obtained are recorded in the following table (table
3).
Table 3. Phytochemical screening
Extracts
Terpenes Phenolic compounds Flavonoïds Acide phenolic Saponins
Cymbopogon citratus
+
+
+
+
+
Lippia chevalieri
+
+
+
Combretum micratum
+
+
+
+
+
Chrysantellum americanum
+
+
+
+
+
Tetrapleura tetraptera
+
+
+
+
The phytochemical screening is carried out on TLC plates
using specific reagents of some chemical groups. We use
organic extracts obtained by fractionation of aqueous extracts.
The screening of the different extracts revealed the presence
of chemical interest groups in the selective extracts, namely
terpenes, flavonoids, saponins and and phenolic acid. These
results are consistent with the previous report on the
phytochemistry of these different traditional drugs [9, 13, 23].
Preclinical studies have shown that certain compounds such
as flavonoids, phenolic acids, saponins and terpenes, which
have anxiolytic effects in a wide range of animal models, are
involved in the treatment of mental disorders [22]. The
presence of these compounds in all extracts justifies their
sedative and anxiolytic properties. Phenolic compounds,
particularly flavonoids, are found in all the extracts that were
analyzed (figure 2). The content of these secondary
metabolites could be used as an indicator for a quantitative
evaluation of the bioactive compounds.
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3.5 Antiradical properties
The figure below shows the presence of light spots on a
purple background. This shows that the extracts contain
phytocompounds that can trap free radicals [20]. By
superimposing the chromatographic profiles of the aqueous
extract and those of the antiradical activity (figure 3), it is
possible to establish the correspondence between the active
areas and the nature of the compounds present. Indeed, the
antiradical activity is more pronounced in extracts of
Combretum micratum, Lippia chevalieri, Tetrapleura
tetraptera compared to extracts of Cymbopogon citratus,
Chrysantellum americanum, this could be explained by the
high content of flavonoids in these decocted extracts. The
antiradical activity of the different extracts shows that these
extracts have antioxidant properties. The antioxidant activity
of the compounds present in the extracts could also justify
their use in the treatment of several diseases, including mental
illnesses [9, 24].
1 (Cymbopogon citratus, Extracts c); 2 (Chrysantellum americanum,
Extracts c); 3 (Combretum micratum, Extracts c); 4 (Lippia
chevalieri, Extracts c); 5 (Tetrapleura tetraptera, Extracts c); 6
(Cymbopogon citratus, Extracts d); 7 (Chrysantellum americanum,
Extracts d); 8 (Combretum micratum, Extracts d); 9 (Lippia
chevalieri, Extracts d); 10 (Tetrapleura tetraptera, Extracts d).
Fig 2: Highlighting of flavonoids in organic extracts (c and d).
3.4 Phenolic Compounds determination
(1): Cymbopogon citratus; (2): Chrysantellum
americanum; (3): Combretum micratum (4):
Lippia chevalieri; (5): Tetrapleura tetraptera
Table 4: Total phenolic et flavonoids content
Extracts
Cymbopogon citratus
Chrysantellum americanum
Combretum micratum
Lippia chevalieri
Tetrapleura tetraptera
Phenolic
content
186,53 ± 3,90
167,65 ± 2,87
395,38 ± 7,05
202,61 ± 2,50
191,11 ± 1,44
Flavonoïds
content
55,53 ± 0,05
60,78 ± 0,09
105,87 ± 0,05
38,13 ± 0,13
31,19 ± 0,05
Fig 3: Antiradical activity of aqueous extracts
A quantitative phytochemical analysis of the aqueous extracts
shows that the phenolic and flavonoid contents are high in all
extracts. A comparison of the total phenolic and flavonoid
contents based respectively on gallic acid and rutin as
reference compounds indicates that the lowest content of
phenolic compound is obtained with Chrysantellum
americanum (167.65 ± 2.87 mg GAE/g extract) and the
highest content is obtained with Combretum micratum extract
(395.38 ± 7.05 mg GAE/g extract). The aqueous extract of
Combretum micratum also gives the high content of total
flavonoids (105.87 ± 0.05 mg Ru E/g extract).Thus, in the
extract of Combretum micratum the content of total phenolics
and total flavonoids is significantly higher than in the other
extracts.
4. Conclusion
In this study, we analyzed the phytochemical profile of five
plants involved in the treatment of mental disorders including
Cymbopogon
citratus,
Chrysantellum
americanum,
Combretum micratum, Lippia chevalieri and Tetrapleura
tetraptera. It appears from the analysis of the chemical profile
of the different extracts that these plants contain bioactive
compounds such as terpenes, flavonoids, phenolic acids and
saponins. Phenolic compounds, particularly flavonoids, are
present in high concentrations in the aqueous extracts. The
antiradical activity on TLC plate is positive for all the extracts
tested. This shows that these extracts also have an antioxidant
properties. Among the extracts studied, Combretum
micrantum showed the highest content of total phenolic and
total flavonoids. The extract of this plant is also the most
positive in the antiradical test. A combination of these plants
containing Combretum micrantum could prove beneficial and
more effective than any of the five plants used separately.
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